Preparation of a Micropatterned Substrate to Study Schwann Cell Phenotypes

Take a silicon wafer with micropatterns of the desired geometry to grow cells in a defined shape.

Pour a polymer mixture on it and allow it to solidify.

Cut a square stamp of the polymer.

Coat the stamp's patterned surface with a protein layer, which facilitates cell attachment.

Using an air stream, remove unbounded proteins.

Take a polymer-coated coverslip.

Place the patterned side of the stamp on it.

Press the stamp to transfer the pattern. Remove the stamp.

Transfer the coverslip into a well containing a surfactant to coat the bare surfaces. This increases surface hydrophobicity, preventing unwanted cell adhesion.

Remove the surfactant and wash with a phosphate buffer.

Add the Schwann cells and allow them to attach to the micropattern.

Wash with a buffer to remove unattached cells.

Add culture medium and incubate.

As the cells grow along the surface, they take the micropattern shape, enabling a study of Schwann cell phenotype.

To prepare a micropatterned substrate, place a patterned silicon wafer inside a circular 150-millimeter diameter by 15-millimeter height Petri dish and pour degassed PDMS onto the wafer. Solidify the PDMS in an oven at 60 degrees Celsius overnight. Allow t